Difference between revisions of "Part:BBa K2324015"

Revision as of 22:34, 1 November 2017

Arabinose inducible fim operon

This part contains the fim operon under the control of an inducible arabinose promoter. The operon contains six Fim proteins, including FimA, FimI, FimC, FimD, FimF and FimG. When co-transformed with FimH constructs from our project, which have rhamnose-inducible promoters, modified type I pili may be produced. The production of the operon must be initiated by inducing the culture overnight at 0.6 OD at 2% arabinose.

Previous iGEM team, Harvard 2015 has synthesised a very similar part, however we used the modular cloning strategy, which has produced scar-sites at various sequences and it has also been codon optimised by the IDT E. coli codon optimisation software.

Results

We transformed this part into E. coli Top10 to image it using a negative staining method on a TEM to obtain an electron micrograph to visualise pili.

Figure 1 Top10 with the fim operon under the control of promoter P_Ara showed no visible signs of pili expression with insertion of the operon alone.

The same part was then transformed into E. coli ΔFimB to also image it using a negative staining method on a TEM to obtain an electron micrograph to visualise pili.

Figure 2 We transformed a plasmid containing the fim operon under the control of the P_Ara promoter into ΔFimB. It exhibits flagella on their cell surface, but the electron micrograph also shows a suggestion of pili.

Results

This suggests that the P_Ara construct was successful in synthesising pili, without the co-transformation with a FimH construct.

Sequence and Features